Human Dystroglycan Antibody

Catalog # Availability Size / Price Qty
AF6868
AF6868-SP
Detection of Human Dystroglycan by Western Blot.
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Product Details
Citations (4)
FAQs
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Human Dystroglycan Antibody Summary

Species Reactivity
Human
Specificity
Detects human Dystroglycan in direct ELISAs and Western blots.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant human Dystroglycan
Gln28-Val749
Accession # Q14118
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Immunohistochemistry
5-15 µg/mL
See below
Knockout Validated
Dystroglycan is specifically detected in HEK293T human embryonic kidney parental cell line but is not detectable in Dystroglycan knockout HEK293T cell line.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

Western Blot Detection of Human Dystroglycan antibody by Western Blot. View Larger

Detection of Human Dystroglycan by Western Blot. Western blot shows lysates of MCF-7 human breast cancer cell line, SH-SY5Y human neuroblastoma cell line, human muscle tissue, and human placenta tissue. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Dystroglycan Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6868) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for a-Dystroglycan at approximately 100-160 kDa (as indicated) and beta -Dystroglycan at approximately 42-44 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry Dystroglycan antibody in Human Skeletal Muscle by Immunohistochemistry (IHC-P). View Larger

Dystroglycan in Human Skeletal Muscle. Dystroglycan was detected in immersion fixed paraffin-embedded sections of human skeletal muscle using Sheep Anti-Human Dystroglycan Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6868) at 3 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to basement membrane. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Knockout Validated Western Blot Shows Human Dystroglycan Antibody Specificity by Using Knockout Cell Line. View Larger

Western Blot Shows Human Dystroglycan Specificity by Using Knockout Cell Line. Western blot shows lysates of HEK293T human embryonic kidney parental cell line and Dystroglycan knockout HEK293T cell line (KO). PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Dystroglycan Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6868) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for a--Dystroglycan at approximately 110 kDa -and beta -Dystroglycan at approximately 42 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Dystroglycan

Dystroglycan, also DAG-1 (Dystrophin-associated glycoprotein 1) is a 180-200 kDa heterodimeric adhesion molecule that links the cell cytoskeleton to the extracellular matrix. It is found on skeletal muscle, cardiac muscle, fibroblasts, smooth muscle and keratinocytes. DAG-1 binds multiple matrix molecules, including laminin-1 and -2, agrin, and perlecan. Intracellularly, the cytoplasmic tail of DAG-1 contributes to a large 400 kDa complex that interacts with the cytoskeleton. The human DAG-1 preprocursor is a type I transmembrane protein 895 amino acids (aa) in length. It contains a 27 aa signal sequence plus an 868 aa proform that undergoes autocatalysis to generate a 626 aa alpha -chain (aa 28-653), and a 242 aa beta -chain. Mature DAG-1 is a heterodimer composed of noncovalently linked alpha ‑ and beta ‑chains. The alpha -chain possesses one potential Ig-like domain (aa 64-162), a mucin-like region (aa 316-485), and a peptidase S72 domain (aa 500-733). It is O‑glycosylated and runs from 100-160 kDa in SDS-PAGE. The beta -chain is N-glycosylated and runs at 42-44 kDa in SDS-Page. It possesses a short 95 aa extracellular region (aa 654-749) plus a 120 aa cytoplasmic domain (aa 776-895). Membrane cleavage of the beta -chain causes dissociation of the heterodimer and generates a 30 kDa truncated form. Over aa 28-749, human DAG-1 shares 93% aa identity with mouse DAG-1.

Long Name
Dystrophin-associated Glycoprotein 1
Entrez Gene IDs
1605 (Human); 13138 (Mouse); 114489 (Rat)
Alternate Names
A3a; AGRNR; DAG1; Dag-1; DAG156DAG; dystroglycan 1 (dystrophin-associated glycoprotein 1); Dystroglycan; Dystrophin-associated glycoprotein 1

Product Datasheets

Citations for Human Dystroglycan Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

4 Citations: Showing 1 - 4
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  1. Ribitol enhances matriglycan of &alpha-dystroglycan in breast cancer cells without affecting cell growth
    Authors: PJ Lu, JD Tucker, EK Branch, F Guo, AR Blaeser, QL Lu
    Sci Rep, 2020;10(1):4935.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  2. biAb Mediated Restoration of the Linkage between Dystroglycan and Laminin-211 as a Therapeutic Approach for alpha-Dystroglycanopathies
    Authors: N Gumlaw, LM Sevigny, H Zhao, Z Luo, DS Bangari, E Masterjohn, Y Chen, B McDonald, M Magnay, T Travaline, T Yoshida-Mo, W Fan, D Reczek, JE Stefano, H Qiu, C Beil, C Lange, E Rao, M Lukason, E Barry, WH Brondyk, Y Zhu, SH Cheng
    Mol. Ther., 2019;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
    Applications: Western Blot
  3. Ribitol restores functionally glycosylated ?-dystroglycan and improves muscle function in dystrophic FKRP-mutant mice
    Authors: MP Cataldi, P Lu, A Blaeser, QL Lu
    Nat Commun, 2018;9(1):3448.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  4. Structural basis of laminin binding to the LARGE glycans on dystroglycan
    Nat Chem Biol, 2016;0(0):.
    Species: Rabbit
    Sample Types: Protein
    Applications: Western Blot

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